Human papillomavirus (abbreviated as HPV) is one closed circular deoxyribonucleotide virus, of which 118 kinds of subtypes have been found, in which 5, 6, 8, 11, 16, 18, 31, 33 and etc genotypes of virus subtypes are associated with human malignancy. Viral oncogenes (such as E6 and E7) carrying thereof will integrate into host genome along with viral DNA during the early development of carcinoma, which results in expression changes of these two oncogenes (such as E6 and E7). Proteins p53 and pRB respectively encoded by E6 and E7 accelerate degradation of host cell, which greatly contributes to carcinogenicity of HR-HPV. Mechanism of replication and separation during chromosome mitosis interfered by such two oncoproteins, induces sever instability of chromosome. It has been verified by researches that integration of high risk HPV DNA relates to changes of chromosome amount and structure. Mice experiment verifies that papilloma virus induce tumorigensis by disrupting DNA sequence not only through viral oncogene but also via integrating host genome. Therefore, integration of HR-HPV results in enhancements of cell immortalization, uncontrollable proliferation and cell instability. Studying on integration of HPV viral gene is benefit to determine pathologic evolution of infection cells by clinical doctors, predicting that whether the infection cells produce viral damage and apoptosis in general or cacinogenesis step by step. In addition, determination of the integration site is also benefit to re drug target research.
Traditional methods of determining HPV genotype use chip genotyping method and mass spectrometry genotyping, which can only determine HPV genotype, other than HPV integration site.
So far, traditional methods detect viral integration site using target or signal amplification method. However, these methods only detect one kind of HPV genotype one time, or have other technical disadvantages, for example, signal detection problem of hybridization in situ and severity conditions of PCR analysis, which are both disadvantage to detect complex integration. The above traditional methods not only are time-consuming and laborious, but also have difficulties in finding new integration sites.
Therefore, currently the method of determining an HPV integration site in a genome of a human tissue sample still needs to be improved.